Toxicity of Heavy Metals and Biological Defense

Principles and Applications in Bioinorganic Chemistry-VII

Ei-lchiro Ochiai
Department of Chemistry, Juniata College, Huntingdon, PA 16652
Some biologically essential elements have been dis-
cussed in the previous articles of this series (1-6).The pre-
sent article discusses some adverse effects of metallic ele-
ments, particularly those of the so-called heavy metals.
Heavy metals a r e ill-defined hut here a r e considered
mainly cadmium, lead, and mercury. Copper, zinc, and sil-
ver also will be mentioned.

Abundance and Toxicity

A certain degree of correlation was shown to exist be-
tween the abundance in the environment and the level of
elements present i n a biological tissue in a previous article
(3).I t is to he noted (see Figs. 2 and 3 of (3))that the essen-
tial elements tend to be those that are relatively abundant
in the environment. The organisms must have tried to util-
ize the elements that they inadvertently ingested from the
environment. If the elements turned out to he useful to the
organisms, they would have become essential to the organ-
isms. Therefore, there is a better chance for a n element to
become essential, if i t is present abundantly and in a read-
O 1 AtOGght
Background Level

ily accesihle form in the environment (the first basic rule of Figure 1. The excess burden of several elements on the environ-
bioselection ( 1 ) ) . mentlbiosphere; the blackened elements are considered to pose the
greatest dangers; the shaded ones are likely to pose dangers. See
I t must he pointed out, though, that some elements such the text for the meaning of the excess burden value.
as Se and Co have become essential to organisms i n trace
amounts despite their low abundances in the environment.
No other more ahundant element could have substituted
for these elements. On the contrary, a n ahundant element
would not become essential, if i t is not useful to organims.
"Aluminum" may be such a n element. Aluminum is uhiqui-
tous i n the environment, though not necessarily i n a read-
ily accesible form.
How about the elements that organisms would encoun-
ter only rarely? They are the elements that are present a t
low levels on the earth or in inaccessible forms. The heavy
metals such a s lead, mercury, and cadmium belong to this
group. Because most organisms have not had experience to
deal with these elements, the organisms could he subjected
to their adverse effects, ifthey ingested them. Some organ-
isms, however, might have had enough experience to deal
with the adverse effects of these elements over the long
history of their evolutions. If that i s the case, those organ-
isms must have developed some defense mechanisms
against the adverse effects of those elements. Otherwise
they would not have survived. In this sense, the elements
that are rare on the earth tend to he toxic to the organisms Fiaure
" 2,Ionic radii: +Il=divalentcations in hioh-soin
- . state (black cir-
c e, or in on-sp n slate (&'I Ie c IC el, 4 1 ='wa en! cat ons n n gn-
in general. This is only a rough generalization. There are SP n Slate oacn sqmre, or n loa-so n stalc ,bnllc sq-are,
many exceptions, a s mentioned above. That is, some ele-
ments that are rare can become essential to the organisms,
if they are critical to the functioning of the organisms. peared, human activities have interfered with the natural
activities. We have collected, hunted, cultivated, and had
Natural versus Anthropogenic Discharge of Elements the pastures grazed by animals, and, a s a result, ravaged
into the Environment our environments. Since the beginning of the civilization
Nature discharges a large quantity of a variety of the and particularly since the industrial revolution we have
elements into the environment. The biosphere and its in- been extracting the natural rcioul-ccr, utilizing them, and
habitants collectively and/or individually have managed to - - at lcast simx uortion of the utililed matcrlal
discharpln~
cope with the natural discharge of elements. Otherwise the into the environments, in such a way that they might in-
organisms would not have survived. Since mankind ap- terfere with the activities of the nature.

Volume 72 Number 6 June 1995 479

Figure 3. Radii of anions and oxyanions.

Nature discharges elements through geological activi-

ties, weathering, and vulcanism. Their magnitude has
been estimated. I t is more difficult to estimate the anthro-
pogenic discharge level. One such estimate has been made
by Nriagu (7). The ratio of "the total discharge (natural
and antbropogenic) over the natural discharge" can then Figure 4. Log Kvalues: a = ethylenediamin complex formation, b =
be computed. The ratio of "1.0" means no extraneous an- oxalate complex formation, c = complex formation with
NHzCHzCHzSH, d = complex formation with HSCHGHzN-
thropogenic discharge; i.e., it represents the background (CHZCOOH)~, e = Kspfor sulfide, f = Itp
for phosphate.
level. The ratio higher than "1.0" means some extraneous
discharge due to the mankind's activities and can be con-
sidered to be the "extra burden" on the nature and the bio- I t is apparent that Hg(I1) has a n inordinately strong af-
sphere. The ratio or extra burden values are shown in Fig- finity toward SZ. The heavy metal cations Hg(I1) and
ure 1. The figure suggests that we would have serious Pb(I1) are considered to be "soft": whereas. most of the
environmental problems not only with cadmium, lead, and transition metal divalent catlons listed (Fig. 2-41 are con-
mercury, but also with tin, selenium, molybdenum, and, sidered to bc on the borderline between tht. "hard" and the
perhaps, copper, arsenic, chromium, and nickel. Iron and "soft". A "soft" acid prefers to bind to a softer base such as
zinc, though high in the ratio figure, may be tolerated bet- S2-. Hg(I1) is one of the softest cations. The soft metal can
ter by the organisms, because they are essential and used also form a stable o-bonded organometallic compounds.
by organisms in substantial quantities and, as a result, the Typical examples are Hg(CH3I2,Hg(CH3)+and Pb(C2H&.,
organisms have better control mechanisms of the levels for Pb(CzHd3'.
these elements in their bodies.
The situation mentioned above is that of'the average on Selectivity in the Uptake of Elements
thc global scale. It is ~ m s i b l ethat the local concmtration
of acertain element ;auld be much higher than Figure 1 The environmental problem due to the extra burden on
indicates. If so, the environmental problem there could be the biosphere would not arise, if the organisms have effl-
much worse than the figure suggests. cient means to cope with the intrusion of excess quantities
of the elements. There are two levels in the coping mecha-
Chemical and Physical Characteristics of Heavy Metal nisms. The first level of protection is to ingest or take up
Cations the elements discriminately, so a s not to take up unneces-
The ionic radii of some metal cations of interest are sary elements andlor unnecessary quantities. The second
shown i n Figure 2. The heavy metals are significantly level of protection or defense is to render the adverse ef-
larger than the more common cations. A n exception is fects of the elements minimal when the organisms have
Ca(II), whose size is similar to those of Cd(II), Hg(II), and ingested them. Some mechanisms of the latter type will be
Pb(I1). The variation in ionic radius among common tran- discussed later. In this section we will look a t the selectiv-
sition metal cations is relatively small; i.e., within 5- ity in the uptake mechanisms of elements.
10%. Metallic elements can be taken up either a s a cation or
Some metallic elements also can exist often in the oxy- a s a n oxyanion. Exceptions for this general rule include
anion forms. Figure 3 shows the ionic radii of more com- the absorption of a metal-chelate a s a whole, as in the case
mon oxyanions. The variation in size among them is even of Fe(II1)-siderophore(see below). Cell membranes are vir-
smaller than that among cations. The variation among tually impermeable to metallic cations and oxyanions. Up-
similar oxyanions is less than 5%. take of a n element, or any substance for that matter, ulti-
Another important character relevant to the the biologi- mately depends on its binding to a biomolecule (acceptor,
cal behaviors would be the binding to ligands. Ametal cat- carrier protein etc).
ion prefers a type of ligand over another. Figure 4 shows Cations and oxyanions are rather featureless. The only
the stability constants or solubility products for several in- distinguishing factors that organisms can take advantage
terestingligands. The solubility product may not necessar- of are
ily reflect the binding strength, but it has a lot to do with (a)the electric charge,
it. Pb(I1) forms a very insoluble phosphate salt. The major
contributor to this fact is the size of Pb(I1). Its large size ib) the size,
matches that of PO4> better than the other cations (ra- ic) the preferred ligands, and
dius-ratio effect). id) the preferable coordination structure.

480 Journal of Chemical Education

 I n contrast, organic compounds may be easier to distin- Another important factor is a kinetic one. Asubstitution-
guish, as they have distinct structures, including the chi- inert ion such a s Cr(III1, once bound to a biomolecule, can-
rality. not be displaced readily by another ion. This not only
Because the variation in size among oxyanions is rela- would lead to a high selectivity, but also it hinders the sub-
tively small, anions of the same geometrical structure and seauent reaction. which mav reauire release of the ion
of the same electric charge are difficult to distinguish. I t from the biomolecule. ~ubstftutickinertcations, Cr(III),
can be inferred that the hysiological receptorlabsorption Co(II1)(in low-spin), Ru(I1) and Rh(II1) are not used widely
2
site for the essential SO4 may take up such ions a s Se04'- in biological systems. Exceptions, however, include low-
and Moo4% (8)and also that the regular uptake mecha- spin Fe(I1) complexes, vitamin Bl2 coenzyme (Co(III)),and,
nisms for Pod3- could absorb V04%and AsO~~-, too (9).If perhaps, Cr(II1) used to maintain a specific structure of
a n absorption mechanism depends on a chemical reaction, DNA.
rather than a mere binding, a more effective discrimina- The selectivity in cation-uptake, thus, is not very great
tion may be accomplished. But no such mechanism is in general, a s far as a single step-uptake is concerned. The
known except for B02-, which can be bound to a carbohy- cation uptake in single-celled organisms, therefore, may
drate moiety; then the B02-sugar moiety is taken up. not be very discriminatory. The selectivity can be improved
in multicellular oreanisms. because a metal ion must eo
The variation among typical divalent cations is not very through several barriers before i t reaches a target c&.
large either. Thus, the selectivity in uptake cannot be ex- Suppose that two metal ions MI and M2 compete for the
pected very high, if it is based on the size and the electric same binding site a t each step, and that the discrimination
charge alone. I n fact, not many efficient systems have been factor (K,/KJ is 10-to-one a t each s t e and
~ that there are
developed in the organisms to discriminate many of the three ba&iek to go through. Then the overall discrimina-
common divalent cations through the size difference. This tion factor would have become 1000-to-one a t the target
does not necessarily mean that ions of different sizes do cells.
not behave differently. They do, but the difference may not A step or two in the process, however, often utilizes fac-
be sufficiently large to be utilized for discrimination in up- tors other t h a n thermodynamic ones. For example, a
take. chemical reaction may be used, which enhances the selec-
I t needs to be pointed out, however, that organisms do tivity enormously. For example, Fe(I1) and Co(I1) can be
have a developed group of compounds that discriminate absorbed by an active transport mechanism in the intes-
cations such a s Na(I), K(I), Ca(II1, and Mg(I1) according to tine. When they come out to the portal vein side, they need
size and electric charge. These compounds called "ionopho- to be oxidized in order to be bound to the transport protein,
res" (10) have, in general, cyclic structures, which have transferrin. This process is catalyzed by a copper-enzyme,
holes that match closely with the sizes of cations. Some ferroxidase; Co(I1) cannot be oxidized to Co(II1) and hence,
organisms, fungi and bacteria, produce and secrete Fe(II1)- cannot be bound to transferrin. Thus, a t this stage, the co-
specific chelating agents called "siderophores" (11).The en- balt would be left behind and would not be carried to the
tire chelates, rather than Fe(II1) ion, are then taken up by bone marrow or the liver. Therefore, the overall iron selec-
the organisms. There are also ion-specific channels that tivity over cobalt would be enhanced greatly. There is a
discriminate ions by their sizes. For example, a Na(1)- verv. s~ecific
. mechanism for the transoort of cobalt-con-
channel hardly allows K(1) ions to go through. These are ta~ningv~raminB12(cobalamins~
exceptional and very selective cases. Thr di.xussions nbove a ~ ~tol the v cases of metallic ions
There are a t least two different iron-uptake mecha- (aquo species). Some elements, particularly the heavy met-
nisms in the human intestine. One of them absorbs Fe(I1) als. can readilv form stable orpanometallic com~ounds.
actively. I t has been demonstrated that Co(I1) or Mn(I1) FO; example, mercury can form dimethyl mer&ry or
competes for this Fe(I1)-absorption site (12). The selectiv- monomethyl mercury, (CH3I2Hg o r CH3Hg(OOCCH3).
ity of this site is not very great, most likely because Co(I1) These compounds can go through cell membranes more
and Mn(I1) have sizes similar to that of Fe(II), and thus readily than divalent aquo-cations, because of their affin-
can bind the Fe(I1)-binding site, though with different af- ity to the membrane. Such a compound can be taken up
finities. That is, this uptake mechanism cannot discrimi- almost indiscrimimately.
nate Fe(I1) against Co(I1) and Mn(I1) very effectively. This
Toxlcities of Heavy Metals
situation appears to be quite general a s far a s cation ah-
sorptions are concerned. The symptoms of the toxic effects of heavy metals may
vary widely a t the physiological level, but the basic toxicity
Cd(II), Hg(II), and Ph(I1) could very likely enter a cell mechanisms a t the molecular level may be limited. The
through the mechanisms to absorb the essential cations toxicities of heavy metals may be caused by the following
such as Ca(II), because of their similarity in size and elec- mechanisms:
tric charge. These heavy metal cations, however, may not
be able to enter efficiently through the Fe(I1) or other es- 1. Blocking the essential functional groups of biamalecules
sential smaller cation-absorption sites, for their sizes are such as enzymes: Specific amino acid residues, such as
sufficiently different. serine-OH, eysteine-SH and histidine-N often constitute
the active sites of enzymes. Hg(II), for example, binds
Other factors, i.e., "preferred coordination structure" strongly cysteine-SH's,blocking an enzymatic activity.
and "preferred coordinating ligand atom", may be utilized 2. Displacing essential metal ions from biomolecules: Amet-
to enhance selectivity in the cation binding. For example, a1 ion mav disolace a native ion. if its afinitv to the bind-
the earlier transition metal ions prefer "O-atom" over "N-
atom" as the ligand, while the later transition metal ions
tend to bind "N-atom'' ligands better, and the heavy metal
cations bind with sulfur strongly. Unfortunately, these fac- eially enzymes and perhaps polynucleatides: A coordina-
tors are rather subtle and have not been utilized fully by tion of a cation may change the conformationof a protein,
rendering it nan-functional.
organims to enhance the selectivity. It might be pointed 4. Disrupting the integrity of biomembranes: Ametal cation
out, though, that the strong affinity toward sulfhydryl may bind the negatively-charged head(s1of phospholipids
groups is employed by a special protein, metallothionein, and the integral protein residues of the membrane.
to store or control the level of heavy metals including cad- 5. Modifying some other biologically active agents: For ex-
mium, mercury, copper, and zinc (see below). ample Cd(I1)and PMII) appear to potentiate the endotax-

Volume 72 Number 6 June 1995 481

 ins produced by bacteria (13).This might be due to their Biological Defenses against Heavy Metals
effect to .block
.-. some enzymes which degrade the endo- When confronted with the toxic effects of heavv metals.
toxm, as m ( 1 ) . the organisms had had to develop some means to defend
6. Binding with bioanions, resulting in a decreased level of
themselves against them, As mentioned earlier, not all or-
essential bioanions, especially P043-or a displacement of
an essential from biaminerals:F~~example,P~(II), ganisms have encountered all the heavy metals equally.
having a size similar to that of Ca(II),could replace Ca(I1) The that were the metals did
in a bone mineral. AS a result the mechanical strength of have developed mechanisms to combat them. 0 t h organ-
the
. .hone mav
~~~
" be affected.The size and the electric charee
~~~~
~~~~~~~~~~ ~
isms mav not have had enoueh .. ex~erience
. to deal with the
\rould he a n irnpurtnor fnrtor in rhese rfrects.One ufthe heavy metal.;, and thus have only relatively inefTecti\x?
basic toxic rffect.i ut I'b 11, is considered 01t w bmding uf mtt:tns to cornhat them. The extraneous anthro~ogen~c dls-
PO,,&, rendering its cytoplasmic level very low. charge of the heavy metals into the environment a s out-
lined above thus poses a serious problem. We will briefly
These toxicity mechanisms are all based on the strong explore some of the biological defense mechanims against
binding abilities of these metallic ions. And a s in the case heavy metals. m i s is not meant to be an exhaustive sur-
of selectivity in uptake mechanism, substitution of a me- vey, but i t is given to illustrate some chemical p,.inciples
tallic ion by another is relatively simple. This is so except behind the biological defense mechanisms,
for substitution-inert metals. In practice, however, any
metal cation bound to a large, somewhat rigid hiomolecule General Tolerance
is kinetically difficult to displace, though not necessarily
substitution-inert. This does not necessarily apply to the Most elements seem to he tolerated by organisms to cer-
cases of proteins of a rather flexible conformation, as in the tain limited degrees. The degree of tolerance is highly de-
case of metallothionein discussed later on. What happens pendent on the organisms, their life stages, a s well as on
often is that a replacement takes place an apo.pro. the elements. This is rather non-specific. Metal ions are to
k i n hinds a cation. A wrong cation can he incorporated in be confined to locales that have little influence on the gen-
., .
this stem if vresent there. era1 biological activities. Such places include cell wall,
some noncritical portions of the cell membranes, vacuoles,
Cd(I1) and Hg(II), for example, can replace the native lysosomes (191, hard tissues such as bone, and extra body
Zn(I1) ion from many proteins and enzymes to a degree tissues such a s hark and hair. As long as the metallic ions
that depends on their affinities. Cu(I), M I ) and Au(I) also are confined in these non-critical places, they may not ex-
have similar characters to Zn(II1, Cd(II), and Hg(I1) be- hibit their toxic effects. Obviously the extent of tolerance
cause of their iso-electronicity. I t would he needless to by these means is limited and highly dependent on the
point out that other cations such a s Cu(II), Ni(II), etc. also structures of the organsims. The toxic symptoms will
may displace a native cation such a s Zn(I1) from enzymes manifest when a metal ion level exceeds a certain thresh-
and proteins. For example, the carcinogenicity of Ni(I1) is old level, unless other more specific defense mechanisms
considered to he due to its replacement of Zn(I1) and/or are available.
Me(I1). . "
". ., the essential cations in DNA oolvmerase. Because
of its different size, Ni(I1) seems to increase the chance of secretion
o f ~ u c u sMaterial
binding wrong nucleotides, thus resulting in the formation
of DNA of a wrong sequence (15). This is found especially in fish. Mucus consists of anionic
Many symptons of Cd(II) toxicity are believed to arise polysaccharides, such a s chondroitin sulfate, ascophyllan,
from the displacement of Zn(II) by Cd(I1). An example is and fucoidan. These contain carhoxylate and/or sulfate
the testicular necrosis by Cd(I1). Testes produce sperms. groups, and they bind cations, It has been that
Hence, the DNA polymerase activity is quite high. DNA fish killed by a high level of Pb(I1) or Zn(I1) were found to
polymerase is a Zn(I1) enzyme, whose activity can be jeop- have high levels of mucus around their gills (20). I t has
ardized by substitution by Cd(I1). (See ref. (14) for a list of suggested that mucus may function as a defense
enzymes affected by Cd(I1)). mechanism against cations.
H d I I ) shows a n enormous affinity toward sulfide a s Metallothioneins
seenrn Figure 4. Because the cystei&c sulfhydryl group
play a rntulytic role in a number of mzymvs, their eruy- Metallothioneins (MT) are low molecular weight (6-
matic actlvltles are susceptihlr to IlgrII,. Or the binding of 70001, cysteine-rich proteins t h a t bind metals such a s
Hg\Il, to amino acid residurs can change a protein contbr- Zn(II), Cd(II), and Hg(I1) (23). MT and its analogues are
mation sufficiently to render it inactive. Kxamples of en- widely distributed among organisms, from bacteria and
zvmes suscc:ptible to Hg 11, include NR/K-A'~I'as(: (the 50- fungi to plants and mammals. A typical mammalian (Zn,
called s o d i i m pump); alkaline phosphatase, lactate Cd) MT is folded into two domains that form separate
dehydrogenase, and glucose-6-phosphatase (14). polynuclear metal cysteine thiolate clusters; Cd4-Cysll
cluster in the C-terminal domain and Zn3-Cyss cluster in
Ph(I1) also inhibits a variety of enzymes. However, the the N-terminal domain. This structure has been deter-
most interesting is its inactivating effects on the enzymes mined hv X-rav crvstalloera~hv(21). and a m e a r s to be
involved in the heme synthesis (16). Particularly impor- commonto most MTS whiih h i d seven metaiiic ions. Me-
tant are ALA dehydratase (ALA-D) and ferrochelatase tallic ions that form M7-MT include Co(I1). Ni(I1). HdII).
(FC). [ALA= baminolewlinic acid]. ALA-D, also called Ph(II), and Bi(II1) in addition to Zn(I1) and Cd(11)'i22):
"porphohilinogen synthase", is a pivotal enzyme in the Ag(1) and CuU) can displace Zn(I1)or Cd(I1)from a MT and
heme synthesis, and FC catalyzes the insertion of Fe(I1) bind in the form of MIS-MT(22). Hg(I1) then displaces 12
into protoporpyrin. The inhibition of ALA-D results in a n Ag(1) or Cu(1) ions and form Hg7-MT (22).
increased level of ALA in urine, which is considered to be a It has been demonstrated that synthesis of MT can be
sensitive measure of lead poisoning. The level of the non- induced by CdUI), Zn(II), Hg(II), and &(I) (23). This fact
fuctioning metal-free or Zn(I1)-porphyrin will increase indicates, a s suggested hy many researchers, that MT
when FC is inhibited. I t also has been reported that the could function a s a protective means against toxic ele-
glohin protein synthesis is affected by lead (17). ments such as Cd(II), Hg(II), and M I ) . Zn(I1) is an essen-
The toxicities of h e a w metals have been reviewed ex- tial element (3). and MT also is involved in the control of
tensively in references 14 and 18. Zn-metabolism. The chemical basis of these functions is

482 Journal of Chemical Education

obviously the strong affinity of these ions toward thiolate. shown to inhibit Cd-induced destruction of nonowlating
'Ibxic metals, thus, bound to MT cannot exhibit their ad- ovaries and placenta, and has protected against the terato-
verse effects. However, lowering pH of the medium or genic effects of cadmium (30). I t is interesting to note that
other yet-to-be-identified mechanisms can release the met- the retention of Cd by tissues actually is increased in the
al ions from MT, and hence, MT may control their cytosolic presence of selenium. A similar effect has been observed in
levels, and also may function a s a temporary storage for
certain elements such a s Zn(I1).
-
reeard to selenium versus mercurv (30). Tuna and bonito
tend to accumu1:itr mercury to a high level: vet the fish
The metabolism of MT is not very well understood. How- thtmst!lve.i art: h t ~ l t h vThe tuna =,as found to contain also
ever, it has been reported that Cd-MT is degraded rapidly a high level of selenLm, and the HgISe mole ratio was
in kidney cells, releasing Cd(II), which is considered to be close to one-to-one (31).Subseauentlv. a linear relations hi^
the cause of the proximal tubular damage (24). The situ- also was reported for Hg versus ~e'contentsin seals and
ation in liver seems to be different. For exam~le.
. . the .
Dres- other marine mammals (32) a s well a s mercury-mine
ence of MT decreases significantly the release of metals workers (33).
--
(Cu and Zn) into bile.. suggesting- that the metals remain Lead Inclusion Body
chelated to MT.
Metallothioneinlike Compounds A dense granular structure (lead inclusion body=LIB)
was first discovered in the nuclei of renal tubular lining
The descriptions above apply to the mammalian type of cells and hepatic cells of children dying of acute lead poi-
MTs. Other organisms produce MT-like proteins that are soning (34). LIB is observed a t smaller lead dosages, even
not exactlv the same a s the mammalian MT. For examole. before a n increased excretion of ALA ensues (see above).
a yeast copperthionein can be induced by Cu(II), but noi b i Similar inclusion bodies have since been reported in other
-
Zn(I1)or HdII). I t is distinct from the mammalian MT. but
the 12 cysteine residues are conserved. The protein pro-
animals (34) and also a moss (35). The LIB isolated from
the kidneys of poisoned rats contained a n average of 57
tects cells against comer uoisoning. but is auuarentlv
.. dis- m m (fresh basis) of lead. The whole kiduevs of the uoi-
pensable foqnormal ielluiar grow& (25). &ned rats contained 0.8 ppm, while the levil of Pb inAthe
Manv authors had r e ~ o r t e dthat Pb(I1) did not induce control kidnevs was 0.009 m m . The LIB consists of a dense
MT, but Ikebuchi et al.in 1986 reported that a MT-like central core and an outer Kbrillar zone. The protein of the
protein containing Pb(I1) was induced along with a Zn(I1)- LIB has a high content of aspartic acid, glutamic acid, gly-
MT in the rat liver when lead acetate was administered cine, cysteine, and tryptophan. The lead in LIB can be re-
(26). It appeared to be very similar to the Zn-MT(II), but moved readily in vitro by EDTA. I t has been suggested that
distinct in electrophoresis. It still is not clear whether the the formation of LIB serves as a protective mechanism
Pb-MT was induced by Pb(I1) rather than Zn(I1) which against lead poisoning (34).
may have contaminated the lead acetate sample, though
its zinc content was reported to be less than 0.05%. The Conversion to Readily Excretable Forms
imuortance of a MT-like rote in in detoxifvin~ . ., Pb(I1) has A single-celled organism is small in size, and hence, if a
not been well estnhlinhrd, though PO 11, does tnnd lo a pre-
chemical entity can be changed to a volatile form, it can
fmmvd MT a s nientloned nbow Pbt 11, mav not induce the
readily diffuse out of the cell. Hg(I1) can be converted into
synthesis of MT, not because it cannot bind to MT or a fac-
tor to induce its synthesis, but rather because Pb(I1) may Hg(0) (36) or (CH&Hg (37)in some bacteria. Both of these
be trapped by phosphate entities present in vivo before it forms of mercury are much less toxic t h a n Hg(I1) or
CH3Hgt, a s well as they have significant vapor pressures.
reaches the MT synthetic a .~.o a r a t u s .
Methylcobalamin has been demonstrated to effect the
Cadmium-resistant Pseudomonos putido produces
methvlation of Sn(I1. IV) and Pb(I1. N)a s well a s He(I1)
three Cd!Il,-binding proteins: 3815,7216, and 7239 in mo-
lecular weight (271.-fhey contain six to eight cysteine resi-
dues and bind cadmium, zinc, and copper. Coordinating li-
fense mecLanism, a s ? c H ~ ) ~ A is~reahily
+ excretable
gands seem to be cysteiue residues,-histidine nitrogen, through the kidneys (9).
and, perhaps, glutamate carboxylates.
Suspension cultures of Rauuolfia serpentina (a higher Concluding Remarks
plant) produced heavy-metal complexing peptides when
grown in a medium containing CdS04 (28). The peptides It has been shown that the toxic effects of and the bio-
were identified to be (y-glutamate-cysteine),-glycine(n=2 logical defenses against heavy metals can be understood
to 11). The composition suggests that the peptides are the basicallv in terms of the fundamental chemical urinci~les
derivatives of glutathione. The peptides, called "phyto- and tbebasic properties of these elements. I t also has been
chelatins". were shown to be induced also bv Cu(I1). HdII). armed that the toxicities of rare elements are caused uar-
Pb(II), and Zn(I1). The induction of the peptides b i C ~ I I ~ tially by the fact that the organisms rarely have encoun-
also was observed in cell cultures of other ~ l a n t including
s
Bereris stolonifera, Fumaria paluiflora, and Galium mol-
-
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